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Barbra and Johnny visit their father's grave in a remote cemetery when they are suddenly set upon by zombies. Barbra manages to get away and takes refuge in what seems to be an abandoned farm house. She is soon joined by Ben who stopped at the house in need of gas. Beset by the walking dead all around them Ben does his best to secure the doors and windows. The news reports are grim however with creatures returning to life everywhere. A group of people hide from bloodthirsty zombies in a farmhouse. Barbra and Ben are surprised when they realize there are 5 people hiding out in the basement: Harry, Helen and Judy Cooper; and a young couple, Tom and Judy. Dissensions sets in almost immediately with Harry Cooper wanting to be in charge. As their situation deteriorates, their chances of surviving the night lessen minute by minute.

During production it was known as Night of Anubis and Night of the Flesh Eaters. The film stars Duane Jones as Ben and Judith O'Dea as Barbra. The plot revolves around the mysterious Corporeal reanimation of the dead and the efforts of Ben, Barbra and five others to survive the night while trapped in a rural Pennsylvania farmhouse.

George Romero produced the film on the small budget of $114,000, but after a decade of theatrical re-releases it had grossed an estimated $12 million in the United States and $30 million internationally. Night of the Living Dead was strongly criticized at the time of its release for its graphic content, but three decades later the Library of Congress entered it into the United States National Film Registry with other films deemed "historically, culturally or aesthetically important."

The culture of Vietnam-era America had a tremendous impact on the film. It is so thoroughly laden with critiques of late-1960s American society that one historian described the film as "subversive on many levels." While not the first zombie film made, Night of the Living Dead influenced countless films and is perhaps the defining influence on the modern pop-culture zombie archetype. The film is the first of five Dead films (completed or pending) directed by Romero. It has been remade twice, in 1990 and in 2006.